Introduction

Abandoned mine drainage (AMD) is the result of abandoned mining operations. AMD is high in sulfuric acid, heavy metals, and metalloids. Treatment of AMD can be carried out via passive remediation systems. Passive remediation systems consist of large settling ponds that promote the biogeochemical precipitation of AMD contaminants. Since these systems are open to the environment, they are naturally colonized by microbial populations. Microbes within these populations metabolically impact the remediation process, both positively and negatively. Isolate AV1 from the Boyce park passive remediation system pond 1 has been identified to positively impact iron remediation and negatively impact manganese remediation. Characterization of this microbial isolate is important to understand more about this novel isolate and understand how it can impact AMD remediation. Characterization of isolate AV1 includes understanding how the isolate grows under different conditions. This is facilitated via growing AV1 in rich media such as R2A as well as minimal media M63.

Methods

Isolate AV1 was grown in the shaker at 30C with either R2A pH 4.0 or M63 media. M63 minimal media was supplemented with an additional carbon source, either lactate or acetate. All conditions were grown at 30C. Different subculture ratios were used for the rich media to mirror those that had been manually obtained in the lab. Data for this experiment was collected using a 96-well plate and plate reader. Controls for the experiments included sterile R2A pH 4.0 and M63 supplemented with either lactate or acetate to be used as a blank and remove background noise.

Results

Growth Condition 1: AV1 R2A pH 4.0 30C 1:25 subculture


#### Condition 1 Individual Replicates ### Condition 1 Average & Standard Deviation of Replicates ### Condition 1 Exponential Growth

Growth Condition 2 AV1 in R2A pH 4.0 30C 1:50 subculture

###Condition 2 Individual Replicates ### Condition 2 Average & Standard Deviation of Replicates ### Condition 2 Exponential Growth

Growth Condition 3

###Condition 3 Individual Replicates

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 2 rows containing missing values (geom_point).

### Condition 3 Average & Standard Deviation of Replicates ### Condition 3 Exponential Growth

Growth Condition 4

###Condition 4 Individual Replicates

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 6 rows containing missing values (geom_point).
## Warning: Removed 2 rows containing missing values (geom_point).

### Condition 4 Average & Standard Deviation of Replicates ### Condition 4 Exponential Growth

Growth Condition 5

###Condition 5 Individual Replicates

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 7 rows containing missing values (geom_point).
## Warning: Removed 5 rows containing missing values (geom_point).

### Condition 5 Average & Standard Deviation of Replicates

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis

### Condition 5 Exponential Growth

Growth Condition 6

###Condition 6 Individual Replicates ### Condition 6 Average & Standard Deviation of Replicates ### Condition 6 Exponential Growth

Growth Condition 7

###Condition 7 Individual Replicates

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 35 rows containing missing values (geom_point).

### Condition 7 Average & Standard Deviation of Replicates ### Condition 7 Exponential Growth

Eight replicates in one plot

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 2 rows containing missing values (geom_point).

Doubling Time for All Conditions

Discussion

Eight replicates in one plot

## Warning in self$trans$transform(x): NaNs produced
## Warning: Transformation introduced infinite values in continuous y-axis
## Warning: Removed 2 rows containing missing values (geom_point).

## Warning in as_grob.default(plot): Cannot convert object of class
## rowwise_dftbl_dftbldata.frame into a grob.

Doubling Time for All Conditions

Discussion